• Asian Pacific Journal of Cancer Prevention
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• v.16 no.14
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• pp.5635-5641
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• 2015

Background: Cirrhosis is regarded as a possible end stage of many liver diseases, including viral infection. It occurs when healthy liver tissue becomes damaged and is replaced by scar tissue and finally may lead to hepatocellular carcinoma. Interferons (IFNs)are two general categories, type I and II. Type I includes one beta interferon and over 20 different alpha interferons. Alpha interferons are very similar in how they work, interacting with other proteins on cells like receptors. The main objective of this study was to compare Mx gene productivity post different cell line treatment with imported and Egyptian biosimilar locally produced IFNs, as well as the efficacy of those tested IFNs. Also, an assessment was made of sensitivity of different cell lines as alternatives to that recommended for evaluation of antiviral activity. Materials and Methods: Different cell lines (Vero, MDBK and Wish) were employed to evaluate cytotoxicity using the MTT assay. Antiviral activity was evaluated compared with standard IFN against VSV, Indiana strain -156, on tested rh-IFNs (imported; innovated and Egyptian biosimilar locally produced IFNs) in the pre-treated cell lines previously mentioned. The virus was propagated in the Wish cell line as recommended. Finally we estimated up-regulation of the Mx gene as a biomarker. Results: Data recorded revealed that test IFNs were safe in test cell lines. Viability was around 100%. Locally tested interferon did not realize the international potency limits, while the imported one was accepted compared with the standard IFN. These results were the same either using infectivity titer reduction assay or crystal violet staining of residual non- infected cells. Mx protein production was cell type related and confirmed by the detected Mx gene expressed in imported and locally produced IFN pre-treated cell lines. The expression of the gene was arranged in the order of Vero> wish > MDBK for the imported IFN, while for the Egyptian biosimillar locally produced one it was MDBK> Vero> wish. With regard to the antiviral activity there was a significant difference of imported IFN potency compared with the locally produced IFN (P<0.05), the IFN potential (antiviral activity) was not cell line related and showed non-significant difference for each separate product. Conclusions: Vero cells can be used as an alternative cell line for evaluation of IFN potency in case of unavailable USP recommended cell lines. Alternative potency evaluation assay could be used and proved significant difference in IFN potency in case of local and imported agents. Evaluation of antiviral activity could be used in parallel to viral infectivity reduction assay for better accuracy. Mx gene can be used as a marker for IFN potential.

• Asian-Australasian Journal of Animal Sciences
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• v.31 no.8
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• pp.1366-1372
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• 2018

Objective: A disintegrin and metallopeptidase with thrombospondin motifs type 8 (ADAMTS8) is crucial for diverse physiological processes, such as inflammation, tissue morphogenesis, and tumorigenesis. The chicken ADAMTS8 (chADAMTS8) gene was differentially expressed in the kidney following exposure to different calcium concentrations, suggesting a pathological role of this protein in metabolic diseases. We aimed to examine the molecular characteristics of chADAMTS8 and analyze the gene-expression differences in response to toll-like receptor 3 (TLR3) stimulation. Methods: The ADAMTS8 mRNA and amino acid sequences of various species (chicken, duck, cow, mouse, rat, human, chimpanzee, pig, and horse) were retrieved from the Ensembl database and subjected to bioinformatics analyses. Reverse-transcription polymerase chain reaction (RT-PCR) and quantitative PCR (qPCR) experiments were performed with various chicken tissues and the chicken fibroblast DF-1 cell line, which was stimulated with polyinosinic-polycytidylic acid (poly[I:C]; a TLR3 ligand). Results: The chADAMTS8 gene was predicted to contain three thrombospondin type 1 (TSP1) domains, whose amino acid sequences shared homology among the different species, whereas sequences outside the TSP1 domains (especially the amino-terminal region) were very dif­ferent. Phylogenetic analysis revealed that chADAMTS8 is evolutionarily clustered in the same clade with that of the duck. chADAMTS8 mRNA was broadly expressed in chicken tissues, and the expression was significantly up-regulated in the DF-1 cells in response to poly(I:C) stimulation (p<0.05). These results showed that chADAMTS8 may be a target gene for TLR3 signaling. Conclusion: In this report, the genetic information of chADAMTS8 gene, its expression in chicken tissues, and chicken DF-1 cells under the stimulation of TLR3 were shown. The result suggests that chADAMTS8 expression may be induced by viral infection and correlated with TLR3-mediated signaling pathway. Further study of the function of chADAMTS8 during TLR3-dependent inflammation (which represents RNA viral infection) is needed and it will also be important to examine the molecular mechanisms during different regulation, depending on innate immune receptor activation.

• Fisheries and Aquatic Sciences
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• v.11 no.4
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• pp.195-204
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• 2008

Viral diseases are major emerging problems of shrimp that have affected the production, and even complete losses for shrimp farms. In this study, we developed a sensitive TaqMan real-time PCR method to quantify white spot syndrome virus (WSSV) and hepatopancreatic parvovirus (HPV) in the shrimp and pond water in which fleshy shrimp, Fenneropenaeus chinensis, and Pacific white shrimp, Litopenaeus vannamei, are reared. WSSV and HPV in pond seawaters ranged from $1.65{\times}10^3$ to $2.43{\times}10^9$ and from 0 to $4.43{\times}10^5$ copies/L of seawater, respectively. Of 20 ponds analyzed, all pond water and shrimp were positive for WSSv. L. vannamei showed higher susceptibility to WSSV than F chinensis. HPV was detected only in the pond water for F chinensis. In shrimp tissue, however, HPV was found in both species, with 23-times higher infection rate in F chinensis than L. vannamei. The total bacterial counts in the pond water ranged from $2.23{\times}l0^3$ to $1.98{\times}l0^5\;CFU/mL$. The variations in total bacterial count for each pond appeared to correlate to the variations of the WSSV load. Statistical analysis indicated that there was no significant difference (P>0.05) between the WSSV load in pond water and shrimp, and there was no relationship between total bacterial load and viral load in the pond water. However, a significant difference (P<0.01) was found between HPV load and L. vannamei and F chinensis pond water.

• The Plant Pathology Journal
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• v.33 no.4
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• pp.393-401
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• 2017

Efforts to control viral diseases in crop production include several types of physical or chemical treatments; antiviral extracts of a number of plants have also been examined to inhibit plant viral infection. However, treatments utilizing naturally selected microorganisms with activity against plant viruses are poorly documented. Here we report isolation of a soil inhabiting bacterium, Pseudomonas oleovorans strain KBPF-004 (developmental code KNF2016) which showed antiviral activity against mechanical transmission of tobamoviruses. Antiviral activity was also evaluated in seed transmission of two tobamoviruses, Pepper mild mottle virus (PMMoV) and Cucumber green mottle mosaic virus (CGMMV), by treatment of seed collected from infected pepper and watermelon, respectively. Pepper and watermelon seeds were treated with culture supernatant of P. oleovorans strain KBPF-004 or control strain ATCC 8062 before planting. Seeds germinated after treatment with water or ATCC 8062 yielded about 60% CGMMV or PMMoV positive plants, whereas < 20% of KBPF-004-treated seeds were virus-infected, a significantly reduced seed transmission rate. Furthermore, supernatant of P. oleovorans strain KBPF-004 remodeled aggregation of PMMoV 126 kDa protein and subcellular localization of movement protein in Nicotiana benthamiana, diminishing aggregation of the 126 kDa protein and essentially abolishing association of the movement protein with the microtubule network. In leaves agroinfiltrated with constructs expressing the coat protein (CP) of either PMMoV or CGMMV, less full-size CP was detected in the presence of supernatant of P. oleovorans strain KBPF-004. These changes may contribute to the antiviral effects of P. oleovorans strain KBPF-004.

• Journal of Society of Preventive Korean Medicine
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• v.16 no.3
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• pp.139-153
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• 2012

Objectives : NK cells are spontaneously cytotoxic lymphocytes. These are not only important parts in the first line of defence against bacterial and viral infections of outside, but they may also play a critical role in chronic viral diseases. NK cells kill their targets spontaneously, without the need for prior sensitization and class I MHC restriction by the regulation of cytolytic functions and secretion of a variety of cytokines, such as interleukin-12(IL-12), MCP-1, IL-6, TNF-${\alpha}$, IFN-${\gamma}$. In addition, macrophage and NK cells cooperate through the production of cell mediates. These cooperation and modulation are one of major factors to prevent for evading immune surveillance of cancer. Hence, it could be assumed that if any candidate to enhance activities of macrophage and NK cell, it is considered as a potentially useful agents against cancer. Methods : In our study, to investigate effect of fermented soybean extracts by Lactic acid bacteria (SFE, soybean fermented extracts) work on intestinal immune cell to maintain general immune modulating and anti-cancer activity. We analyzed NK cytotoxicity assay and gene expressions of cytokine related with macrophage and NK cell activity. Results : In vitro experiment, SFE was verified as safety material for cell toxicicty to tumor cell strain without any toxicity of tumor growth inhibition and various cell strain. Effects of macrophage activity stimulating directly by SFE measured induced cytokine. The studies showed that IL-12 production by stimulation of SFE depended on concentration from 0.16mg/mL to 0.63mg/mL with non toxicity to cell, and it was the best activity at 0.63mg/mL. Besides, the effective concentration of SFE producing TNF-${\alpha}$ is similar to IL-12, but it was the best activity at 1.25mg/mL. The level of MCP-1, IL-6 and IFN-${\gamma}$ depended on concentration from 0.16mg/mL to 10mg/mL, IFN-${\gamma}$ showed the best activity at the effective concentration of 0.63mg/mL. With the result of NK cell activity measurement, the spleen cell of mouse injected SFE had 1.5 times higher killing effect than non injected cell. Conclusions : The result of this studies is that Soybean fermetated extracts(SFE) has possibility to immune aided material for the function not only inhibition of microbial infection to macrophage but also activity of adaption immune and cellular immune system.

• Pediatric Infection and Vaccine
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• v.24 no.2
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• pp.87-94
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• 2017

Purpose: The inappropriate prescription of antibiotics in children with upper respiratory tract infection (URTI) is common. This study evaluated the factors that influence antibiotics use in hospitalized children with viral URTI confirmed by reverse transcriptase-polymerase chain reaction (RTPCR) assay. Methods: The medical records of admitted patients who performed RT-PCR assay for respiratory virus pathogens from January 2013 to November 2014 were examined. The demographic and clinical features were compared between patients who were administered antibiotics at admission and those who were not. We also investigated differences between children who continued antibiotics and those who stopped antibiotics after a viral pathogen was identified. Results: In the total 393 inpatients, the median age was 23 months (interquartile range, 13 to 41.3 months). Antimicrobial agents were prescribed in 79 patients (20.1%) at admission. Patients with acute otitis media (AOM) had higher rates of antibiotics prescription than those without AOM (48.1% vs. 2.2%, P<0.001), with an adjusted odds ratio of 91.1 (95% confidence interval, 30.5 to 271.7). Level of high-sensitivity C-reactive protein and the proportion of acute rhinosinusitis were also significantly associated with antibiotics use (P<0.001). Among the 44 patients with viruses identified using the RT-PCR method during hospitalization, antibiotic use was continued in 28 patients (63.6%). AOM was statistically associated with continued antibiotic use in the patients (P=0.002). Conclusions: Although the respiratory virus responsible for URTI etiology is identified, clinicians might not discontinue antibiotics if AOM is accompanying. Therefore, careful diagnosis and management of AOM could be a strategy to reduce unjustified antibiotic prescriptions for children with URTI.

• The Plant Pathology Journal
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• v.40 no.1
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• pp.73-82
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• 2024

Gardenia (Gardenia jasminoides) is a popular and economically vital plant known for its ornamental and medicinal properties. Despite its widespread cultivation, there has been no documentation of plant viruses on gardenia yet. In the present study, gardenia leaves exhibiting symptoms of plant viral diseases were sampled and sequenced by both metatranscriptome and small RNA sequencing. As a consequence, bean common mosaic virus (BCMV) was identified in gardenia for the first time and named BCMV-gardenia. The full genome sequence of BCMV-gardenia is 10,054 nucleotides (nt) in length (excluding the poly (A) at the 3' termini), encoding a large polyprotein of 3,222 amino acids. Sequence analysis showed that the N-termini of the polyprotein encoded by BCMV-gardenia is less conserved when compared to other BCMV isolates, whereas the C-termini is the most conserved. Maximum likelihood phylogenetic analysis showed that BCMVgardenia was clustered closely with other BCMV isolates identified outside the leguminous plants. Our results indicated that the majority of BCMV-gardenia virus-derived small interfering RNAs (vsiRNAs) were 21 nt and 22 nt, with 21 nt being more abundant. The first nucleotide at the 5' termini of vsiRNAs derived from BCMV-gardenia preferred U and A. The ratio of vsiRNAs derived from sense (51.1%) and antisense (48.9%) strands is approaching, and the distribution of vsiRNAs along the viral genome is generally even, with some hot spots forming in local regions. Our findings could provide new insights into the diversity, evolution, and host expansion of BCMV and contribute to the prevention and treatment of this virus.

• International Journal of Industrial Entomology and Biomaterials
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• v.12 no.2
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• pp.57-61
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• 2006

Suppressor of cytokine signaling (SOCS) is known to be as a negative feedback regulator in Janus kinase signal transducer and activator of transcription signaling. Highly conserved SOCS box domain was cloned from a Korean malaria vector, Anopheles sinensis. Sequence analysis indicates that it has identity to Anopheles gambiae (96%), Aedes aegypti (94%), Drosophila melanogaster (78%), Mus musculus (72%) and Homo sapiens (72%), respectively. Tissue specificity RT-PCR demonstrated that the expression level of AsSOCS transcript was high at abdomen, midgut, and ovary, whereas developmental expression patterns showed that the level of AsSOCS was high at egg, early pupae, and adult female. On the other hand, RT-PCR analysis after bacterial challenge showed that SOCS mRNA was strongly induced in larvae. In addition, it was also induced by various immune elicitors such as lipoteicoic acid, CpG-DNA, and laminarin. It seems that AsSOCS, repressor of JAK-STAT pathway, is highly conserved in mosquito, and may play an important role in mosquito innate immune response.

• The Plant Pathology Journal
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• v.22 no.3
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• pp.239-247
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• 2006

In 2003, a survey of sweet potato virus disease was carried out in seed boxes as well as in various sweet potato fields. Virus infection rate was $5\sim100%$ and 100% at seed boxes and fields, respectively. No relationship of the disease incidence and severity was observed between sweet potato cultivating areas and cultivars. A total of 179 samples were collected and analyzed based on serological, electron microscopic and molecular properties. Field-grown sweet potatoes were examined to inspect 8 different viruses using NCM-ELISA, resulting that 30% of sweet potato was infected by one virus, whereas 70% was by more than 2 viruses. However, RT-PCR using primers selected for seven viruses, such as Sweet potato feathery mottle virus (SPFMV) revealed that of one-hundred seventy-nine tested; 71 of SPFMV, 29 of SPGV, 19 of SPFMV+SPGV, 1 of SPFMV+SwPLV, 1 of SPFMV+SPLCV, 2 of SPFMV+SPGV+SwPLV, 6 of SPFMV+SPGV+SPLCV, 2 of SPFMV+SPGV+SwPLV+SPLCV and 48 of unknown viruses were identified from the field samples. In root, viral diseases were severer in Yeoju than in Mokpo Experiment Station and infection rate was much different depending on sweet potato cultivars.

• Journal of Veterinary Clinics
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• v.30 no.6
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• pp.453-455
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• 2013

Heartworm infection is a fatal disease causing heart failure and pulmonary diseases in dogs. This heartworm infection can also occur in wild carnivores including Raccoon dogs. Recent study found that relatively high prevalence rate in wild Raccoon dog population. Therefore, this study was designed to evaluate the prevalence rate of D. immitis in free-roaming Raccoon dogs and the recovery rate of microfilariae in infected Raccoon dogs in Korea. Overall prevalence rate of D. immitis in Korean Raccoon dogs was 17.8%. Prevalence rate in male Raccoon dogs was 21.8%, while that in female Raccoon dogs was 12.8%. Microfilariae were not detected in 17 Raccoon dogs having positive in heartworm antigen test. Our study result suggested that the prevalence rate of D. immitis in Korea is twice higher than that of Japan. In addition, microfilaremia is rare in Raccoon dogs as commonly noticed in cats.